Ultrastructure of the Lyonet's glands in larvae of Diatraea saccharalis Fabricius (Lepidoptera : Pyralidae)

The Lyonet's gland is found in Lepidoptera larvae, close to the excretory duct of the silk gland. The role played by this gland is still uncertain. This work aims to describe the ultrastructure of the Lyonet's bland in Diatraea saccharalis larvae, offering suggestions regarding its possible function. The insects were reared under laboratory-controlled conditions. The glands were conventionally prepared for transmission (TEM) and scanning (SEM) electron microscopy. SEM showed that Lyonet's glands are paired small structures located in the ventral side of the head. They are composed by clustered long cells resembling leaves. Under TEM observations, each cell is surrounded by a thin basal lamina and contains large stellate nucleus. The cytoplasm presents large and empty canaliculi with small microvilli. The basal plasma membrane forms numerous infoldings where numerous and well-developed mitochondria are concentrated. The cytoplasmic membrane system is poorly developed. Our ultrastructural results suggest that the Lyonet's gland in D. saccharalis larvae may be involved in the uptake of small molecules from the hemolymph no morphological evidences of macromolecules synthesis and secretion were noticed. The detection of nerve fibers in the gland suggest a neural control for the glandular cell function.

SEQUENCE OF A CDNA-ENCODING BOTHROPSTOXIN-I, A MYOTOXIN FROM THE VENOM OF BOTHROPS-JARARACUSSU

With the aim of further understanding the structure/function relationships in the membrane-damaging activity of the Lys(49) phospholipase A(2) (Lys(49)-PLA(2)) sub-family, we used PCR (polymerase chain reaction) on total venom gland cDNAs from Bothrops jararacussu with degenerate oligodeoxyribonucleotides encoding the N- and C-termini of myotoxin II, a Lys(49)-PLA(2) from Bothrops asper. A 350-bp cDNA coding for bothropstoxin I (BtxtxI) was amplified. Sequencing of the amplified fragment shows that BtxtxI has a Lys(49), and comparison with the known structure of myotoxin II showed that the amino acids involved in the formation of a novel dimeric structure in this protein were also conserved.

Reproductive compatibility and chromosome pairing in the Drosophila buzzatii complex

Fertility (percentage of fertile crosses) and the degree of synapsis in salivary gland chromosomes in isofemale lines of Drosophila buzzatii, D. serido, D. koepferae and D. seriema were analysed. D. buzzatii was completely sterile in intercrosses with strains from the other species except for D. koepferae. The other species intercrossed to a greater or lesser degree, but also differed between crossing directions. Homologous pairing in salivary gland preparations of strains and hybrids conformed with the data on fertility. The lowest degree of synapsis was present in hybrids between D. koepferae and D. buzzatii, D. seriema and D. koepferae and D. koepferae and D. serido. These species also exhibited the lowest degree of reproductive compatibility. Hybrids between D. seriema and D. serido showed an intermediate degree of synapsis (pairing absent in the proximal and distal chromosome ends), as well as fertility greater than that found in the other interspecific crosses. Results of the fertility of crosses involving strains of a single species, compared with data in the literature, indicated that intraspecific divergence occurred in D. serido and D. koepferae.

Anxiety levels and wild running susceptibility in rats: assessment with elevated plus maze test and predator odor exposure

It is reported in the literature that nearly 20% of rats are susceptible to displays of wild running (WR) behavior when submitted to high intensity acoustic stimulation. Some characteristics of WR suggest that it can be viewed as a panic-like reaction. This work aimed to test whether WR-sensitive rats show higher levels of anxiety in elevated-plus-maze (EPM) and predator-odor exposure paradigms in comparison with WR-resistant ones. Male adult Wistar rats were submitted to two trials of acoustic stimulation (104 dB, 60 s) in order to assess WR susceptibility. Seven WR-sensitive and 15 WR-resistant rats were evaluated by the EPM test. Other 13 WR-sensitive and 18 WR-resistant animals were submitted to the predator-odor exposure test which consisted of a 10 min-session of free exploration in a specific apparatus containing two odoriferous stimuli: cotton swab imbedded with snake cloacal gland secretion or with iguana feces (control). WR-sensitive rats presented a significantly higher closed-to open-ann-entry ratio in the EPM test. All rats responded with anxiety-like behaviors to the predator odor exposure, although the WR-sensitive ones showed a marked behavioral inhibition regardless of the odor condition. We conclude that WR-sensitive rats present elevated levels of anxiety manifested by means of passive behavioral strategies. (C) 2004 Elsevier B.V. All rights reserved.

Vascular organization of the hystricomorph placenta: a comparative study in the Agouti, capybara, guinea pig, paca and rock cavy

The placental vasculature of five hystricomorph rodents was examined by latex injection of the blood vessels, immunohistochemistry and scanning electron microscopy of vessel casts. The pattern of branching of the vessels is described at the level of fine structure. The placenta is divided into lobes separated by interlobular trophoblast. Fetal arteries course through the interlobular areas and give rise to capillaries from which blood drains into veins at the centre of the lobes. Maternal blood reaches the placenta through spiral arteries that pass around the perimeter of the subplacenta. They supply large maternal blood sinuses, lined by trophoblast, which run through the interlobular areas and into the centre of the lobes. Here they supply fine channels that run parallel to the fetal capillaries, so that maternal blood flows from the centre of the lobe to the periphery. This arrangement provides the morphological basis for countercurrent exchange. The maternal channels of the labyrinth drain into spaces formed by the latticework of the interlobular trophoblast and thence through venous lacunae to a basal venous lacunar ring. The subplacenta is supplied by a single fetal artery. The vessels within the subplacenta pursue a tortuous course with dilatations and constrictions as in an endocrine gland. (C) 2003 Elsevier Ltd. All rights reserved.

SCANNING ELECTRON-MICROSCOPY OF THE RHODNIUS-NEGLECTUS (HEMIPTERA) LABIAL SALIVARY-GLANDS AFTER STARVATION

Labial salivary glands are found in the majority of insects. They are relatively large, extend back into the thorax, and in Rhodnius, they are cherry red in color due to a pigment derived from traces of hemoglobin absorbed form the gut. In most insects they are acinous shaped, with long excretion channels that present differentiated regions which from salivary reservoirs. The glands may be relatively simple or complexly branched and convoluted. In Rhodnius they are described as being unilobed with no traces of division. The main duct leaves the gland at its anterior extremity. The acini have different kinds of cells but all of them are seen as sources of secretion. Our material has a different shape due to the fact that the animals spent 20 days under starvation conditions. New data are also obtained through treatment with collagenase and HCl. The importance of the study of these glands lies in the fact that it will further understanding of the transmission of Chagas' disease.

Electrophoretical studies of proteins of the hypopharyngeal glands and of the larval food of Melipona quadrifasciata anthidioides Lep (Hymenoptera, Meliponinae)

The electrophoretical protein patterns of hypopharyngeal glands, larval food of Melipona, and royal jelly of Apis were compared.Since protein patterns of hypopharyngeal glands from newly emerged workers, brood cell provisioners and foragers are similar to freshly deposited larval food, the identical protein bands probably represent actual gland secretion. This suggests that, as in Apis, the glands secrete proteins to the larval food, and maintain this ability throughout life, although at slightly different intensities, according to the activity of the bees.The similarity on the electrophoretic profiles of the major larval food protein in Apis and Melipona is an interesting finding because of its probable evolutionary significance.

Partial genetic characterization of Stearoyl Coa-Desaturase's structural region in Bubalus bubalis

Conjugated Linoleic Acids (CLAs) comprise a family of positional and geometric isomers of linoleic acid. The main form of CLA, cis-9, trans-11-C18:2 show positive effects in cancer prevention and treatment. The major dietary sources of these fatty acids are derived from ruminant animals, in particular dairy products. In these animals, the endogenous synthesis mainly occurs in mammary gland by the action of enzyme Stearoyl CoA Desaturase (SCD). Different levels of expression and activity of SCD in mammary gland can explain partially the variation of CLA levels in fat milk. Considering a great fat concentration in bubaline milk and the benefit of a high and positive correlation between fat milk and CLA production, this study was carried on with the intention of sequencing and characterizing part of the gene that codifies SCD in buffaloes. Genomic DNA was extracted from blood samples of lactating bubaline which begins to the breed Murrah. After the (acho que nao precisa desse the) extractions, PCR (Polymerase Chain Reaction) reactions were made by using primers Z (sic) (sic) D1 and E1 (sic) (sic) F1. The fragments obtained in PCR were cloned into T vectors and transformed in competent cells DH10B line. After this, three samples of each fragment were sequenced from 5' and 3' extremities using a BigDye kit in an automatic sequencer. Sequences were edited in a consensus of each fragment and were submitted to BLAST-n / NCBI for similarity comparisions among other species. The sequence obtained with Z (sic) (sic) D1 primers shows 938 bp enclosing exons 1 and 2 and intron 1. The primers E1 (sic) (sic) F1 show 70 bp corresponding to exon 3 of bubaline SCD gene. Similarities were obtained between 85% and 97% among bubaline sequences and sequences of SCD gene described in human, mouse, rat, swine, bovine, caprine and ovine species. This study has permitted the identification and partial characterization of SCD codifing region in Bubalus bubalis specie.

Use of PCR-RFLP (Polymerase Chain Reaction-Restricted Fragment Length Polymorphism) in the gene of the enzyme Stearoyl-CoA-Desaturase in Bubalus bubalis

The milk is an important food because it contents Conjugated Linoleic Acids (CIA). These fatty acids are synthesized in mammary gland under action of the enzyme Stearoyl CoA-Desaturase (SCD) and have showed some positive effects in human disease prevention and treatments. A variation of CLA in milk fat exists and can be partially explained by the different levels of expression of SCD. The aim was to study part of the encoding regions of SCD's gene using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). Genomic DNA was extracted from lactating Murrah females. After this, PCR reactions were made by using primers Z (sic) (sic) D1 that encloses exon I, II and intron I. The fragments amplified are composed by 938 pb. Then, RFLP techniques were applied in the fragments using the restriction enzymes Pst I and Sma I. The enzyme Pst I has generated fragments of 788pb and 150bp and the Sma I has generated fragments of 693pb and 245pb. All the animals showed the same migration standard for both enzymes, characterizing a genetic monomorphism for this region of SCD gene. The analysis determined that there aren't genetic differences between these animals in the studied regions by using Pst I and Sma I enzymes.

REDUCTION OF CELL PROLIFERATIVE ACTIVITIES OF GASTRIC STUMP ADENOMATOUS HYPERPLASIAS AFTER BILE REFLUX DIVERSION IN RATS

Previously we reported the majority of lesions induced hy bile reflux, in the absence of chemical carcinogens, in the rat remnant stomach to consist primarily of gastric type and secondarily of intestinal type cells, and that they are reversible after diversion of bile reflux. The present study was designed to evaluate changes in proliferative activities in cells of each type under these conditions. The frequency of adenomatous hyperplasia (AH) induced in the gastric stump mucosa by duodenal content reflux after Billroth II partial gastrectomy (BII) increased until the 54th week of the experiment. Roux-en-Y (RY) surgical procedure which prevents duodenal reflux performed at the 24th or 36th week after BII led to a decrease in AH. Cell content of the lesions was analyzed using routine H&E staining, immunohistochemical staining for pepsinogen isoenzyme 1 and histochemical procedures for mucins (paradoxical concanavalin A, galactose oxidase Schiff and sialidase galactose oxidase Schiff reactions) and proliferation in each compartment evaluated by an immunohistochemical method using bromodeoxyuridine (BrdU) and a monoclonal antibody against BrdU. At the 54th week the number of BrdU-labeled cells per normal pyloric column was significantly (P < 0.05) increased to 10.63/pit after the BII operation, while it diminished to 5.23/pit after RY diversion, this being the same level as with the RY procedure alone. AH maintained a high rate of BrdU incorporation at 12.7% after BII operation, which was also significantly reduced (P < 0.01) to 7.0% by the RY surgery. The intestinal type cell showed highest (22.2%), the surface mucous type cell showed the next (16.5%) and the pyloric gland type cell showed lowest (5.2%) BrdU labeling indices after BII operation. All the cell types in AH showed similar proportional decreases in BrdU incorporation after RY diversion. Thus surgical intervention reverses the cell proliferation caused by bile reflux in the gastric stump.

Morphological changes in the salivary glands of Amblyomma cajennense females (Acari : Ixodidae) in different feeding stages on rabbits at first infestation

This study examined salivary glands of unfed, partially engorged, and engorged females of the tick Amblyomma cajennense on rabbits at first infestation using histological and histochemical techniques. In type I acini, no significant changes were observed among the three feeding conditions. In type II acini of unfed females, c1, c2, and c4 cells were described for the first time in this species. In a comparison among the three feeding conditions, an increase in this acinus was observed, due to the increase in secretion in c1, c2, and c4 cells and the appearance of c3 cells. In engorged females, some cells were still active. Type III acini presented cells d, e, and f containing secretion in unfed females. In partially engorged females, these cells were devoid of secretion. In engorged females, type III acini exhibited a reduced lumen. After engorgement, all acini underwent a degenerative process, as observed in females after two to five days post-engorgement.

A NEW SPECIES of THE BOKERMANNOHYLA MARTINSI GROUP FROM CENTRAL BAHIA, BRAZIL WITH COMMENTS on BOKERMANNOHYLA (ANURA: HYLIDAE)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Pediatric Chronic Sclerosing Sialadenitis: Kuttner Tumor

Chronic sclerosing sialadenitis is an uncommon cause of salivary gland enlargement mainly occurring in the fifth and seventh decade of life. In the Western population, chronic sclerosing sialadenitis has been characterized as an IgG4-related disease. Although rare, this lesion occurs in children. To increase awareness about this entity in the pediatric age group, we report the case of an 11-year-old boy with a hard, 4.0-cm circumscribed mass in the right submandibular gland. Histologically there was marked distortion of the gland architecture by a dense lymphocytic infiltrate and extensive fibrosis with septa that crossed and distorted the gland, leaving atrophic acini and dilated, irregular ducts. The lymphoid infiltrate formed multiple follicles with active germinal centers, numerous plasma cells, and areas with diffuse arrangement. Immunophenotyping showed abundant CD20- and CD3-positive lymphocytes; cytokeratin AE1/AE3 highlighted the distorted architectural pattern; IgG staining showed large numbers of positive cells infiltrating the interstitium and surrounding the atrophic acini and ducts. IgG4 staining revealed a large proportion of positive infiltrating elements. Kuttner tumor belongs to the group of IgG4-related sclerosing diseases. The differential diagnosis includes pleomorphic adenoma and other salivary gland neoplasms. Its recognition in children is important clinically because this entity is amenable to steroid treatment, and additional work up and follow up is warranted to stave off other IgG4-related diseases/complications.

Morphological changes in the cephalic salivary glands of females and males of Apis mellifera and Scaptotrigona postica (Hymenoptera, Apidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)